• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

墨脱地区番茄青枯菌的分离鉴定及生防菌剂效果评价

Isolation, Characterization, and Effective Biocontrol Agents of Ralstonia solanacearum Causing Tomato Bacterial Wilt in Medog, Tibet

  • 摘要:
    目的 分离鉴定西藏墨脱番茄青枯病菌,明确其遗传多样性,评价筛选适用于墨脱地区青枯菌的生防菌剂,为当地番茄青枯病防控提供理论和技术支持。
    方法 对采自西藏墨脱地区的番茄青枯病病株通过组织分离法进行病原菌的分离纯化, 通过形态观察结合分子鉴定明确其种类;通过演化型多重PCR(phylotype-specific multiplex polymerase chain reaction, Pmx-PCR)、生理小种和生化型鉴定确定分离菌株的类型;利用PCR扩增获得egl基因部分序列进行系统发育分析,明确分离菌株的序列变种;采用平板抑菌圈法测定5种生防菌剂对病原菌的抑制效果。
    结果 从墨脱地区的6株番茄青枯病病株中分离获得8株病菌,经鉴定为青枯菌(Ralstonia solanacearum),所分离菌株均属于演化型Ⅰ,生化型Ⅲ和序列变种48。室内药剂抑菌结果表明,所测试的5种生防菌剂对墨脱番茄青枯菌均具有良好的抑菌效果,其中昶百润微生物菌剂的效果最好,抑菌直径可达到19 mm以上。
    结论 分离鉴定了西藏墨脱地区的番茄青枯菌菌株,明确了其群体遗传结构;筛选出可适用于墨脱地区的生防菌剂,为青枯病的高效绿色防控提供技术支持。

     

    Abstract:
    Objective The pathogen causing tomato bacterial wilt in Medog, Tibet was isolated, identified, genetic diversity clarified, and effective biocontrol agents determined.
    Method Potential pathogens that caused the bacterial wilt on tomato plants were isolated and purified from the specimens collected in Medog by tissue separation. Specific species were identified based on morphological characteristics and molecular analysis prior to toxicologic classification using the phylotype-specific multiplex PCR (Pmx-PCR), physiological race, and biovar determination. Partial egl sequences were amplified by PCR for phylogenetic analysis to assign sequevars. Inhibitory effects of 5 biocontrol agents against the isolated pathogen were assessed using the agar plate diffusion method (inhibition zone assay).
    Result Eight Ralstonia solanacearum strains were isolated from the sampled 6 tomato plants infected with bacterial wilt in Medog. All isolates belonged to Phylotype I, Biovar III, and Sequevar 48. In vitro inhibition tests on the isolate demonstrated significant efficacies of all 5 agents. Of them, Changbairun Microbial Agent showed the largest average inhibition zone diameter that exceeded 19mm.
    Conclusion R. solanacearum strains from Medog were isolated, characterized, and population genetic structure defined. An effective biocontrol agent was identified for eco-friendly control of the disease on tomato plants.

     

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